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73 SUPERNATANT OF ESTROGEN-TREATED HUMAN UMBILICAL CORD BLOOD DERIVED HEMATOPOIETIC STEM CELLS UNDER EXTREMELY HYPOXIA ATTENUATE EXPERIMENTAL: SPINAL CORD INJURY S Chen 1 , W Lo 2 , H Chang 2 , C Wu 1 1 Chi Mei Medical Center, Tainan, Taiwan, 2 Stem cell Research Center, Health Banks Co., Ltd., Taipei, Taiwan From our previous studies, both of human umbilical cord blood derived hematopoietic stem cells (hUCBHSCs) transplantation and estrogen (E2) administration can improve hindlimb motor dysfunction after experimental spinal cord injury (SCI) respectively. It indicated these two agents may have the potentials of anti-inflammation, vasculogenesis and neurogenesis. However, there is still persisted graft-versus-host disease (GVHD) in hUCBHSCs transplantation therapy. Therefore, we designed 17bestradiol (E2)-stimulated hUCBHSCs cultured in HSC medium under extreme hypoxic (0.1% O 2 ) for three days. Three days later, the conditioned medium (CM) harvested from supernatant (cell free). Rats were divided into six groups: (1) sham operation (laminectomy only); (2) SCI + 0.5 cc saline, iv, N¼6; (3) SCI + 0.5 cc HSC medium, iv, N¼6; and (4) SCI + 0.5 cc CM1 (HSCs-110 6 + HSC medium), iv, N¼6; (5) SCI + 0.5 cc CM2 (E2+HSCs-110 6 + HSC medium), iv, N¼6; and (6) SCI + 0.5 cc CM3 (E2+ HSC medium), iv, N¼6. SCI was induced by compressing the spinal cord (T8-T9) for 1 min with an aneurysm clip cali- brated to a closing pressure of 55 g. All the administered were injected immediately after SCI via the tail vein. Behavioral tests of motor function invented by Basso, Beattie, Bresnahan (BBB) scoring was detected at day 1 to 7 after SCI. The terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assay was also conducted after SCI to evaluate spinal cord apoptosis. The Immunohistochemical staining of glial fibrillary acidic protein was conducted to evaluate astrogliosis. Enzyme-Linked Immunosorbent assay to analyze the amount of vascular endothelial growth factor (VEGF) and glial cell line-derived neurotrophic factor (GDNF) in the CM. It was found systemic administration of estrogen-treated HSC in HSCs medium under 0.1% O 2 significantly attenuated the SCI induced hind limb dysfunction and spinal cord apoptosis. Both GDNF and VEGF could be detected significantly higher in the CM2. 74 WILL NOT BE PRESENTED 75 SUCCESSFUL USE OF PLERIXAFOR IN HARD TO MOBILIZE PATIENTS - FOLLOWING HIGH DOSE THERAPY ALL PATIENTS DEVELOPED FAST AND SUSTAINED EN- GRAFTMENT WITH DURABLE CLINICAL RESPONSES D Josefsen, G Hetland, A Blystad, Y Fløisand, G Kvalheim Oslo University Hospital, Oslo, Norway Mobilization of autologous hematopoietic stem cells is most frequently per- formed using G-CSF alone. In our clinic we use G-CSF in combination with chemotherapy. In 15-20% of the patients insufficient numbers of CD34+ cells are harvested, and are regarded as poor-mobilizers. We have previously shown that mobilized patients appearing with low concentration of CD34+ cells (5-10 cells/mL) and recovered total white cell count >1010 9 /L, can successfully harvest stem cells by addition of plerixafor. Moreover, poor mobilizing lymphoma patients have a less favorable prognosis than good mobilizers. Here, we have mobilized and harvested 33 poor-mobilizing cancer patients by addition of plerixafor. Furthermore, the 16 lymphoma patients were followed up after reinfusion of autologous stem cells with regard to engraftment as well as clinical response. The day prior to harvest of the 33 patients, the level of leukocytes was 21,810 9 cells/L (median), and the CD34+ concentration was 5,510 6 /L (median). Following plerixafor injection, the concentration of CD34+ cells increased to 26,810 6 /L (median). The patients were then successfully har- vested (median: 3,910 6 CD34+cells/kg) with 1-2 days of apheresis. High dose therapy is a curative treatment for lymphomas. Therefore, we focused especially on the clinical outcome on these16 patients. Following high dose therapy, time to short term engraftment, defined by neutrophils >0.510 9 /L and thrombocytes >2010 9 /L were 11 days (median) and 20 days (median). Moreover, in contrast to previous findings we have observed durable responses with relapse free survival of 77% and overall survival of 93%, with a median observation time of 14 months. In conclusion, our findings show that addition of plerixafor in hard to mobilize patients allow them to proceed to high dose therapy. Moreover, the additional costs related to the use of plerixafor can be justified since lymphoma patients obtain similar event free and overall survival as those patients that were good mobilizers. 76 ESTABLISHING AN ALGORITHM TO ENSURE AN OPTIMAL YIELD OF MOBILISED PROGENITOR CELLS PG Dyson 1 , S Hiwase 1 , LB To 1,2 , I Lewis 1,2 1 SA Pathology, Adelaide, Australia, 2 Royal Adelaide Hospital, Adelaide, Australia Aim: Haemopoietic reconstitution post transplantation depends on dose of haemopoietic progenitor cells (HPC) infused. Collection of an adequate dose HPC depends on effective mobilisation of HPC from the marrow into the circulation. Poor mobilisation affects patient outcome and resource utilisation. To maximise HPC harvest we sought to optimise mobilisation and collection protocols by identifying developing an algorithm that would ensure collection of optimal numbers of HPC. Method: We reviewed data for 128 patients who underwent progenitor cell mobilisation and autologous transplantation in our institution in 2009-2011. For this study we defined the transplant CD34 + cell dose as being optimal (610 6 /kg for MM and 310 6 /kg for NHL), low (2 to 610 6 /kg in MM and 2 to 310 6 /kg in NHL) and poor (<210 6 /kg in MM and NHL). Results: The target CD34 + cell dose was achieved during the first mobi- lisation in 100/128 (78%) patients. Multiple mobilisation cycles were per- formed in 19/128 (15%) patients. CD34 + cell yield correlated with circulating pre CD34 + levels - Spearman r 2 ¼0.51. An optimal dose could not be collected in patients who failed to reach CD34 + /ml ¼ 15. Conclusion: In both patient groups a CD34 + /ml < 15 predicted an optimal CD34 + cell dose would not be collected. For G-CSF mobilised patients if the day 5 CD34 + /ml < 7 then collection of an optimal cell yield is unlikely. For chemotherapy ¼ G-CSF mobilized patients if the day 10 white cell count < 2 10 9 /l then collection of an optimal cell dose is unlikely. 77 G-CSF PRIMED DONOR HAEMATOPOIETIC STEM CELL COLLECTIONS ARE ASSOCIATED WITH REDUCED VIABLE T CELL YIELD FOR DONOR LYMPHOCYTE INFUSION V Antonenas 1 , F Garvin 1 , K Yehson 1 , G Hansra 1 , D McCulloch 2 , E Blyth 2 , M Hertzberg 2 , D Gottlieb 1,2 1 Sydney Cellular Therapies Laboratory, Westmead Hospital, Australia, 2 Blood and Marrow Transplant Service, Westmead Hospital, Australia Collection details Details Multiple Myeloma (MM) Lymphoma (NHL) Gender Male 45 31 Female 35 17 Age 60 (24 - 70) 55 (19 - 69) First mobilisation G-CSF 12 2 Chemo + G-CSF 68 46 First mobilisation dose Optimal 66 34 Suboptimal 14 14 Second mobilisation dose Optimal 2 4 Poor 5 2 Low 3 2 No collection - 1 S24 Poster abstracts
Transcript of WILL NOT BE PRESENTED
Collection details
DetailsMultiple
Myeloma (MM)Lymphoma(NHL)
GenderMale 45 31Female 35 17
Age 60 (24 - 70) 55 (19 - 69)First mobilisationG-CSF 12 2Chemo + G-CSF 68 46
First mobilisation doseOptimal 66 34Suboptimal 14 14
Second mobilisation doseOptimal 2 4Poor 5 2Low 3 2No collection - 1
S24 Poster abstracts
73SUPERNATANTOF ESTROGEN-TREATEDHUMAN UMBILICALCORD BLOOD DERIVED HEMATOPOIETIC STEM CELLSUNDER EXTREMELY HYPOXIA ATTENUATE EXPERIMENTAL:SPINAL CORD INJURYS Chen1, W Lo2, H Chang2, C Wu11Chi Mei Medical Center, Tainan, Taiwan, 2Stem cell Research Center,Health Banks Co., Ltd., Taipei, Taiwan
From our previous studies, both of human umbilical cord blood derivedhematopoietic stem cells (hUCBHSCs) transplantation and estrogen (E2)administration can improve hindlimb motor dysfunction after experimentalspinal cord injury (SCI) respectively. It indicated these two agents may have thepotentials of anti-inflammation, vasculogenesis and neurogenesis. However,there is still persisted graft-versus-host disease (GVHD) in hUCBHSCstransplantation therapy. Therefore, we designed 17bestradiol (E2)-stimulatedhUCBHSCs cultured in HSC medium under extreme hypoxic (0.1% O2) forthree days. Three days later, the conditioned medium (CM) harvested fromsupernatant (cell free). Rats were divided into six groups: (1) sham operation(laminectomy only); (2) SCI + 0.5 cc saline, iv, N¼6; (3) SCI + 0.5 cc HSCmedium, iv, N¼6; and (4) SCI + 0.5 cc CM1 (HSCs-1�106+ HSC medium), iv,N¼6; (5) SCI + 0.5 cc CM2 (E2+HSCs-1�106+ HSC medium), iv, N¼6; and(6) SCI + 0.5 cc CM3 (E2+ HSC medium), iv, N¼6. SCI was induced bycompressing the spinal cord (T8-T9) for 1 min with an aneurysm clip cali-brated to a closing pressure of 55 g. All the administered were injectedimmediately after SCI via the tail vein. Behavioral tests of motor functioninvented by Basso, Beattie, Bresnahan (BBB) scoring was detected at day 1 to 7after SCI. The terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphosphate-biotin nick end labeling assay was also conducted after SCI toevaluate spinal cord apoptosis. The Immunohistochemical staining of glialfibrillary acidic protein was conducted to evaluate astrogliosis. Enzyme-LinkedImmunosorbent assay to analyze the amount of vascular endothelial growthfactor (VEGF) and glial cell line-derived neurotrophic factor (GDNF) in theCM. It was found systemic administration of estrogen-treated HSC in HSCsmedium under 0.1% O2 significantly attenuated the SCI induced hind limbdysfunction and spinal cord apoptosis. Both GDNF and VEGF could bedetected significantly higher in the CM2.
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75SUCCESSFUL USE OF PLERIXAFOR IN HARD TO MOBILIZEPATIENTS - FOLLOWING HIGH DOSE THERAPY ALLPATIENTS DEVELOPED FAST AND SUSTAINED EN-GRAFTMENT WITH DURABLE CLINICAL RESPONSESD Josefsen, G Hetland, A Blystad, Y Fløisand, G KvalheimOslo University Hospital, Oslo, Norway
Mobilization of autologous hematopoietic stem cells is most frequently per-formed using G-CSF alone. In our clinic we use G-CSF in combination withchemotherapy. In 15-20% of the patients insufficient numbers of CD34+ cellsare harvested, and are regarded as poor-mobilizers. We have previously shownthat mobilized patients appearing with low concentration of CD34+ cells (5-10cells/mL) and recovered total white cell count >10�109/L, can successfullyharvest stem cells by addition of plerixafor. Moreover, poor mobilizinglymphoma patients have a less favorable prognosis than good mobilizers. Here,we have mobilized and harvested 33 poor-mobilizing cancer patients byaddition of plerixafor. Furthermore, the 16 lymphoma patients were followedup after reinfusion of autologous stem cells with regard to engraftment as wellas clinical response.
The day prior to harvest of the 33 patients, the level of leukocytes was21,8�109 cells/L (median), and the CD34+ concentration was 5,5�106/L(median). Following plerixafor injection, the concentration of CD34+ cellsincreased to 26,8�106/L (median). The patients were then successfully har-vested (median: 3,9�106 CD34+cells/kg) with 1-2 days of apheresis.
High dose therapy is a curative treatment for lymphomas. Therefore, wefocused especially on the clinical outcome on these16 patients. Following highdose therapy, time to short term engraftment, defined by neutrophils>0.5�109/L and thrombocytes >20�109/L were 11 days (median) and 20 days(median). Moreover, in contrast to previous findings we have observed durable
responses with relapse free survival of 77% and overall survival of 93%, witha median observation time of 14 months.
In conclusion, our findings show that addition of plerixafor in hard tomobilize patients allow them to proceed to high dose therapy. Moreover, theadditional costs related to the use of plerixafor can be justified since lymphomapatients obtain similar event free and overall survival as those patients that weregood mobilizers.
76ESTABLISHING AN ALGORITHM TO ENSURE AN OPTIMALYIELD OF MOBILISED PROGENITOR CELLSPG Dyson1, S Hiwase1, LB To1,2, I Lewis1,21SA Pathology, Adelaide, Australia, 2Royal Adelaide Hospital, Adelaide,Australia
Aim: Haemopoietic reconstitution post transplantation depends on dose ofhaemopoietic progenitor cells (HPC) infused. Collection of an adequate doseHPC depends on effective mobilisation of HPC from the marrow into thecirculation. Poor mobilisation affects patient outcome and resource utilisation.To maximise HPC harvest we sought to optimise mobilisation and collectionprotocols by identifying developing an algorithm that would ensure collectionof optimal numbers of HPC.
Method: We reviewed data for 128 patients who underwent progenitor cellmobilisation and autologous transplantation in our institution in 2009-2011.
For this study we defined the transplant CD34+ cell dose as being optimal(�6�106/kg for MM and �3�106/kg for NHL), low (2 to 6�106/kg in MMand 2 to 3�106/kg in NHL) and poor (<2�106/kg in MM and NHL).
Results: The target CD34+ cell dose was achieved during the first mobi-lisation in 100/128 (78%) patients. Multiple mobilisation cycles were per-formed in 19/128 (15%) patients.
CD34+ cell yield correlated with circulating pre CD34+ levels - Spearmanr2 ¼0.51. An optimal dose could not be collected in patients who failed to reachCD34+/ml ¼ 15.
Conclusion: In both patient groups a CD34+/ml < 15 predicted an optimalCD34+ cell dose would not be collected. For G-CSF mobilised patients if theday 5 CD34+/ml < 7 then collection of an optimal cell yield is unlikely. Forchemotherapy ¼ G-CSF mobilized patients if the day 10 white cell count < 2 �109/l then collection of an optimal cell dose is unlikely.
77G-CSF PRIMED DONOR HAEMATOPOIETIC STEM CELLCOLLECTIONS ARE ASSOCIATED WITH REDUCED VIABLE TCELL YIELD FOR DONOR LYMPHOCYTE INFUSIONV Antonenas1, F Garvin1, K Yehson1, G Hansra1, D McCulloch2, E Blyth2,M Hertzberg2, D Gottlieb1,21Sydney Cellular Therapies Laboratory, Westmead Hospital, Australia, 2Bloodand Marrow Transplant Service, Westmead Hospital, Australia
